本文選題：基質(zhì)細(xì)胞衍生因子受體 + 氧化低密度脂蛋白��； 參考：《華中科技大學(xué)》2009年碩士論文

【摘要】：背景基質(zhì)細(xì)胞衍生因子(stromal cell derived-factor1,SDF-1)又名CXCL12或前B刺激因子,屬于CXC族趨化因子成員,同基因編碼兩種蛋白:SDF-1α和SDF-1β。SDF-1主要在骨髓基質(zhì)細(xì)胞及骨髓內(nèi)皮細(xì)胞表達(dá),在造血干細(xì)胞中也有相應(yīng)的表達(dá)。SDF-1唯一的受體CXCR4是一個(gè)有七個(gè)跨膜結(jié)構(gòu)域的G蛋白耦聯(lián)的受體,在造血干/祖細(xì)胞表面及內(nèi)皮祖細(xì)胞表面高表達(dá),新近研究表明CXCR4也在骨髓間充質(zhì)細(xì)胞(marrow stromal cells;MSC)上表達(dá)。文獻(xiàn)報(bào)道,在心肌梗死后的心肌中立即出現(xiàn)了SDF-1的上調(diào)并導(dǎo)致過表達(dá)CXCR4的MSC向損傷心肌遷移。在Zhang等的研究中表明CXCR4+的MSC參與了新生血管的生成并分化為新的心肌細(xì)胞,動(dòng)物實(shí)驗(yàn)表明SDF-1/CXCR4軸的相互作用在心肌修復(fù)時(shí)對(duì)MSC的募集起著重要作用,在SDF-1和/或CXCR4敲除老鼠的研究中表明該軸在細(xì)胞分化中起著重要作用。MSC是骨髓微環(huán)境中重要細(xì)胞成分,具有向各種細(xì)胞分化的潛能,且有文獻(xiàn)報(bào)道MSC可向平滑肌祖細(xì)胞(Smooth muscle progenitor cell;SPC)分化。骨髓源性MSC或SPC是動(dòng)脈粥樣硬化(atherosclerosis,As)斑塊中平滑肌細(xì)胞重要來源之一。那么在As的斑塊形成及修復(fù)過程中SDF-1/CXCR4軸是否能募集MSC或骨髓源性的SPC的歸巢致?lián)p傷動(dòng)脈,參與As形成尚未見報(bào)道。 目的檢測(cè)As危險(xiǎn)因子氧化低密度脂蛋白(Oxidized Low Density Lipoprotein;ox-LDL)對(duì)小鼠骨髓源性SPC SDF-1受體CXCR4表達(dá)的影響,探討SDF-1/CXCR4對(duì)骨髓源性SPC歸巢至受損動(dòng)脈的作用。 方法小鼠骨髓源性SPC與ox-LDL(50μg/mL)共同孵育,用逆轉(zhuǎn)錄聚合酶鏈反應(yīng)法(Reverse transcription-polymerase chian reaction;RT-PCR)檢測(cè)SPC CXCR4 mRNA的表達(dá),免疫印跡法(immunoblotting)及免疫熒光染色和共聚焦激光掃描顯微鏡分析(immunofluorescence staining and confocal laser scanning microscope analysis)檢測(cè)SPC CXCR4蛋白表達(dá),觀察ox-LDL對(duì)SPC表達(dá)CXCR4的時(shí)效關(guān)系。 結(jié)果未給予ox-LDL刺激的SPC有基礎(chǔ)水平的CXCR4表達(dá),50μg/mL的ox-LDL刺激SPC 0-72h,隨刺激時(shí)間的延長(zhǎng),CXCR4表達(dá)逐漸增強(qiáng),36h達(dá)峰值,其mRNA和蛋白水平分別為基礎(chǔ)水平的5.73倍和5.02倍,差異均有統(tǒng)計(jì)學(xué)意義(p0.05),隨后逐漸下降,但仍高于基礎(chǔ)水平。免疫熒光染色和共聚焦激光檢測(cè)并觀察到在0h時(shí)即未給予ox-LDL刺激時(shí)SPC的胞質(zhì)和細(xì)胞膜表面有少量的CXCR4表達(dá),隨著作用時(shí)間的延長(zhǎng),CXCR4的表達(dá)量逐漸上調(diào)且在36h時(shí)其在SPC的胞質(zhì)和細(xì)胞膜表面的表達(dá)最強(qiáng),60h后CXCR4的表達(dá)量逐漸下降。 結(jié)論ox-LDL上調(diào)小鼠骨髓源性SPC CXCR4的表達(dá),提示ox-LDL有可能通過上調(diào)SPC CXCR4的表達(dá)致SPC遷移和歸巢至病變動(dòng)脈參與As形成。
[Abstract]:Stromal cell derived-factor1 (SDF-1), also known as CXCL12 or pre B stimulator, is a member of the CXC chemokines, and the same gene encodes two proteins: SDF-1 A and SDF-1 beta.SDF-1 are mainly expressed in bone marrow stromal cells and bone marrow endothelial cells, and there are also.SDF-1 receptor C in the blood making stem cells. XCR4 is a G protein coupled receptor with seven transmembrane domains, which is highly expressed on the surface of hematopoietic stem / progenitor cells and on the surface of endothelial progenitor cells. Recent studies have shown that CXCR4 is also expressed on bone marrow mesenchymal cells (marrow stromal cells; MSC). It is reported that the up regulation of SDF-1 in the myocardium after myocardial infarction and the result of the overexpression are reported. CXCR4 MSC migrated to the injured myocardium. In the study of Zhang, CXCR4+ MSC participated in the formation of new blood vessels and differentiated into new cardiomyocytes. Animal experiments showed that the interaction of SDF-1/CXCR4 axis played an important role in the recruitment of MSC in the repair of myocardium. In the study of SDF-1 and / or CXCR4 knockout mice, the axis was fine. Cell differentiation plays an important role in.MSC, an important cell component in bone marrow microenvironment, and has the potential to differentiate into various cells. And it is reported that MSC can differentiate into smooth muscle progenitor cells (Smooth muscle progenitor cell; SPC). Myelogenic MSC or SPC is an important source of smooth muscle cells in atherosclerotic (atherosclerosis, As) plaques. First, it is not reported that whether the SDF-1/CXCR4 axis can raise the injured arteries of the MSC or bone marrow derived SPC during the plaque formation and repair of As. The formation of As is not yet reported.
Objective to investigate the effect of As risk factor Oxidized Low Density Lipoprotein (ox-LDL) on the CXCR4 expression of bone marrow derived SPC SDF-1 receptor in mice, and to explore the effect of SDF-1/CXCR4 on the SPC homing of bone marrow to the damaged artery.
Methods the mouse bone marrow derived SPC was incubated with ox-LDL (50 mu g/mL), and the expression of SPC CXCR4 mRNA was detected by reverse transcription polymerase chain reaction (Reverse transcription-polymerase Chian reaction; RT-PCR), and immunoblotting (immunoblotting), immunofluorescence staining and confocal laser scanning microscopy were used to analyze the expression of the expression. Nd confocal laser scanning microscope analysis was used to detect SPC CXCR4 protein expression and to observe the time-dependent relationship between ox-LDL and SPC expression.
The results showed that SPC with no ox-LDL stimulation had the basic level of CXCR4 expression, and the ox-LDL of 50 mu g/mL stimulated SPC 0-72h. With the time of stimulation, the expression of CXCR4 increased gradually and the 36h reached the peak value. The mRNA and protein levels were 5.73 times and 5.02 times of the basic level respectively. The difference was statistically significant (P0.05), and then gradually decreased, but still higher than the base water. Immunofluorescence staining and confocal laser detected and observed that there was a small amount of CXCR4 expression in the cytoplasm of SPC and the surface of the cell membrane in the absence of ox-LDL stimulation at 0h. The expression of CXCR4 increased gradually with the prolongation of action time and the expression of CXCR4 was the strongest in the cytoplasm and surface of the cell membrane at the time of 36h, and the amount of CXCR4 expressed gradually after 60H. Drop.
Conclusion ox-LDL up-regulated the expression of bone marrow derived SPC CXCR4 in mice, suggesting that ox-LDL may be involved in SPC migration and homing to the lesion artery to participate in As formation by up regulation of the expression of SPC CXCR4.

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R329

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本文編號(hào)：1861161